Teilprojekt 3
Caspase-1 ist ein essentieller Regulator von Entzündungsreaktionen durch die proteolytische Prozessierung der Interleukine IL-1β und IL-18. Darüber hinaus konnte kürzlich gezeigt werden, dass Caspase-1 an der unkonventionellen Sekretion seiner Substrate, weiterer Interleukine und Proteine beteiligt ist. Die molekularen Mechanismen und das regulatorische Netzwerk der unkonventionellen ER/Golgi-unabhängigen Proteinsekretion, wie sie in der proinflammatorischen zellulären Immunantwort auftritt, sind noch weitestgehend unbekannt.
Zur Charakterisierung der Sekretome von Zellkulturmodellen für CASP1 Mutationen werden wir eine quantitative Analyse-Plattform entwickeln, die auf hochauflösender Ionenfallen-Orbitrap Massenspektrometrie basiert und auf Isotopenmarkierung verzichtet. Der von uns vorgeschlagene analytische Ansatz soll die unabhängige Durchführung verschiedener Experimente und Kontrollen gestatten und durch die hypothese-freie Analyse zum Verständnis der molekularen Mechanismen beitragen, die der unkonventionellen Sekretion zugrunde liegen.
Teilprojektleiter
Mitarbeiter
Dr. rer. nat. Marc Gentzel
Projektkoordination
Tel.: 0351-210 2614
Email: gentzel@mpi-cbg.de
Mukesh Kumar
Tel.: 0351-210 2647
Email: kumar@mpi-cbg.de
Ausgewählte Publikationen
Groessl M, Luksch H, Rösen-Wolff A, Shevchenko A, Gentzel M. Profiling of the human monocytic cell secretome by quantitative label-free mass spectrometry identifies stimulus-specific cytokines and proinflammatory proteins. J Proteomics 2012; 12(18):2833-42.
Vasilj A, Gentzel M, Ueberham E, Gebhardt R, Shevchenko A. Tissue proteomics by one-dimensional gel electrophoresis combined with label-free protein quantification. J Proteome Res 2012; 11,3680-9.
Schuhmann K, Almeida R, Baumert M, Herzog R, Bornstein SR, Shevchenko A. Shotgun lipidomics on a LTQ Orbitrap mass spectrometer by successive switching between acquisition polarity modes. J Mass Spectrom 2012; 47(1):96-104.
Knaust A, Shevchenko A, Shevchenko A. Horizontal carryover of proteins on one-dimensional polyacrylamide gels may jeopardize gel-enhanced liquid chromatography mass spectrometry proteomic interpretations. Anal Biochem 2012; 421(2):779-81.
Danckwardt S, Gantzert AS, Macher-Goeppinger S, Probst HC, Gentzel M, Wilm M, Gröne HJ, Schirmacher P, Hentze MW, Kulozik AE. p38 MAPK controls prothrombin expression by regulated RNA 3' end processing. Mol Cell 2011; 41(3):298-310.
Spirin V, Shpunt A, Seebacher J, Gentzel M, Shevchenko A, Gygi S, Sunyaev S. Assigning spectrum-specific P-values to protein identifications by mass spectrometry. Bioinformatics 2011; 27(8):1128-34.
Edelson-Averbukh M, Shevchenko A, Pipkorn R, Lehmann WD. Discrimination between peptide O-sulfo- and O-phosphotyrosine residues by negative ion mode electrospray tandem mass spectrometry. J Am Soc Mass Spectrom 2011; 22(12):2256-68.
Krastev DB, Slabicki M, Paszkowski-Rogacz M, Hubner NC, Junqueira M, Shevchenko A, Mann M, Neugebauer KM, Buchholz F. A systematic RNAi synthetic interaction screen reveals a link between p53 and snoRNP assembly. Nat Cell Biol 2011; 13(7):809-18.
Schuhmann K, Herzog R, Schwudke D, Metelmann-Strupat W, Bornstein SR, Shevchenko A. Bottom-up shotgun lipidomics by higher energy collisional dissociation on LTQ Orbitrap mass spectrometers. Anal Chem 2011; 83(14):5480-7.
Schwudke D, Schuhmann K, Herzog R, Bornstein SR, Shevchenko A. Shotgun lipidomics on high resolution mass spectrometers. Cold Spring Harb Perspect Biol 2011; 3(9):a004614. doi:10.1101/cshperspect.a004614. (Review)
Herzog R, Schwudke D, Schuhmann K, Sampaio JL, Bornstein SR, Schroeder M, Shevchenko A. A novel informatics concept for high-throughput shotgun lipidomics based on the molecular fragmentation query language. Genome Biol 2011; 12(1):R8. doi:10.1186/gb-2011-12-1-r8.
Feike AC, Rachor K, Gentzel M, Schambony A. Wnt5a/Ror2-induced upregulation of xPAPC requires xShcA. Biochem Biophys Res Commun 2010; 400(4):500-6.
Thoma C, Fraterman S, Gentzel M, Wilm M, Hentze MW. Translation initiation by the c-myc mRNA internal ribosome entry sequence and the poly(A) tail. RNA 2008; 14(8):1579-89.
Kind J, Vaquerizas JM, Gebhardt P, Gentzel M, Luscombe NM, Bertone P, Akhtar A. Genome-wide analysis reveals MOF as a key regulator of dosage compensation and gene expression in Drosophila. Cell 2008; 133(5):813-28.
Danckwardt S, Kaufmann I, Gentzel M, Foerstner KU, Gantzert A, Neu-Yilik G, Bork P, Keller W, Wilm M, Hentze MW, Kulozik AE. Splicing factors stimulate polyadenylation via USEs at non-canonical 3’ end formation signals. EMBO Jour 2007; 26(11):2658-69.
Niggeweg R, Köcher T, Gentzel M, Buscaino A, Taipale M, Akhtar A, Wilm M. A general precursor ion-like scanning mode on quadrupole-TOF instruments compatible with chromatographic separation. Proteomics 2006; 6(1):41-53.
Gentzel M, Köcher T, Ponnusamy S, Wilm M. Preprocessing of tandem mass spectrometric data to support automatic protein identification. Proteomics 2003; 3(8):1597-610.
Kast J, Gentzel M, Wilm M, Richardson K. Noise filtering techniques for electrospray quadrupole time of flight mass spectra. J Am Soc Mass Spectrom 2003; 14(7):766-76.
Walther TC, Askjaer P, Gentzel M, Habermann A, Griffiths G, Wilm M, Mattaj IW, Hetzer M. RanGTP mediates nuclear pore complex assembly. Nature 2003; 7;424(6949):689-94.
Forler D, Kocher T, Rode M, Gentzel M, Izaurralde E, Wilm M. An efficient protein complex purification method for functional proteomics in higher eukaryotes. Nat Biotechnol 2003; 21:89-92.
Makarov EM, Makarova OV, Urlaub H, Gentzel M, Will CL, Wilm M, Lührmann R. Small nuclear ribonucleoprotein remodeling during catalytic activation of the spliceosome. Science 2002; 298(5601):2205-8.
Groessl M,Zava O, Dyson PJ. Cellular uptake and subcellular distribution of ruthenium-based metallodrugs under clinical investigation versus cisplatin. Metallomics 2011; 3:591-599.
Groessl M, Dyson PJ. Bioanalytical and Biophysical Techniques for the Elucidation of the Mode of Action of Metal-Based Drugs. Curr Top Med Chem 2011; 11(21):2632-46.
Groessl M, Fei Z, Dyson PJ, Katsyuba SA, Vikse KL, McIndoe JS. A mass spectrometric and theoretical study of polyiodides: the connection between solid state, solution and gas phases. Inorg Chem 2011; 50(19):9728-3.
Groessl M, Terenghi M, Casini A, Lobinski R, Dyson PJ. Reactivity of anticancer metallodrugs with serum proteins: new insights from size exclusion chromatography-ICP-MS and ESI-MS. J Anal At Spectrom 2010; 25:305-313.
Groessl M, Tsybin YO, Hartinger ChG, Keppler BK, Dyson PJ. Ruthenium versus Platinum: Interactions of Anticancer Metallodrugs with Duplex Oligonucleotides Characterised by Electrospray Ionisation-Mass Spectrometry. J Biol Inorg Chem 2010; 15:677-688.
Groessl M, Bytzek A, Hartinger ChG. The serum protein binding of pharmacologically active gallium(III) compounds assessed by hyphenated CE-MS techniques. Electrophoresis 2009; 30:2720-2727.
Groessl M, Hartinger ChG, Połeć-Pawlak K, Jarosz M, Dyson PJ, Keppler BK. Elucidation of the Interactions of an Anticancer Ruthenium Complex in Clinical Trials with Biomolecules Utilizing Capillary Electrophoresis Hyphenated to Inductively Coupled Plasma-Mass Spectrometry. Chem Biodivers 2008; 5:1609-14.
Groessl M, Hartinger, ChG, Połeć-Pawlak K, Jarosz M, Keppler BK. CE hyphenated to ICP-MS: a Novel Approach for the Analysis of Anticancer Metallodrugs in Human Serum and Plasma. Electrophoresis 2008; 29:2224-2232.
Groessl M, Hartinger ChG, Dyson PJ, Keppler BK. CZE–ICP-MS as a Tool for Studying the Hydrolysis of Ruthenium Anticancer Drug Candidates and Their Reactivity Towards the DNA Model Compound dGMP. Inorg Biochem 2008; 102:1060-1065.
Groessl M, Reisner E, Hartinger ChG, Timerbaev AR, Arion VB, Keppler BK. Structure–Activity Relationships for NAMI-A Type Complexes (HL)[trans-RuCl4L(S-dmso)ruthenate(III)] (L = Imidazole, Indazole, 1,2,4-Triazole,4-Amino-1,2,4-triazole, and 1-Methyl-1,2,4-triazole): Aquation, Redox Properties, Protein Binding, and Antiproliferative Activity. J Med Chem 2007; 50:2185-2193.